This is a revised application for funding of a DakoCytomation MoFlo high speed fluorescence activated cell sorter. There is currently only one high speed cell sorter available to most investigators at the Albert Einstein College of Medicine (AECOM), which is inadequate to meet current needs. The requested funding will enable us to add a second high speed sorter to our main FACS Core Facility, which will double cell sorting capacity and extend the range of applications that can be supported. The Facility provides most FACS-based resources for approximately 275 users associated with the laboratories of more than 50 NIH- funded investigators at AECOM. The Facility has a highly evolved plan of operations, sound financial oversight, and a long history of strong institutional support. There are two full-time technical staff with extensive background in operation of the MoFlo sorter, and a third person with comparable skills will soon be hired. There is a faculty supervisor (Dr. Steven A. Porcelli, PI of this SIG application) who is a Professor of Microbiology and Immunology and an internationally known research immunologist. A steering committee composed of seven distinguished AECOM faculty meets regularly to set policy for current operations and future growth, and provide quality assurance. The instrument for which we have requested funding will have three lasers to enable excitation of a broad range of different fluorescent probes. Cells and subcellular structures in suspension can be characterized and sorted at speeds exceeding 25,000 events per second using simultaneous measurement of two light scatter parameters and up to seven channels of fluorescence. This instrument permits the isolation of specific cell types from among complex mixed cell suspensions, and is the most powerful and precise method available for purification of cells and certain subcellular structures based on the binding of specific fluorescent probes or antibodies. The requested instrument will be housed in biosafety level-2 (BSL-2) laboratory space and will be equipped with an aerosol containment module, thus enabling us to perform high speed sorting of samples containing infectious agents requiring BSL-2 precautions in accordance with our Institutional Biosafety Committee. The provision of funds to support the purchase of this instrument will accelerate research into the causes and treatment of major public health problems including cancer, diabetes, cardiovascular and infectious diseases. The acquisition of this instrument will also play a significant role in facilitating the continuing growth of AECOM as a major center for advanced biomedical research and education. [unreadable] [unreadable] [unreadable]